Purpose: Biomarkers predicting clinically significant prostate cancer (sPC) before biopsy are currently lacking. This study aimed to develop a non-invasive urine test to predict sPC in at-risk men using urinary metabolomic profiles. Materials and Methods: Urine samples from 934 at-risk subjects and 268 treatment-naïve PC patients were subjected to liquid chromatography/mass spectrophotometry (LC-MS)-based metabolomics profiling using both C18 and hydrophilic interaction liquid chromatography (HILIC) column analyses. Four models were constructed (training cohort [n=647]) and validated (validation cohort [n=344]) for different purposes. Model I differentiates PC from benign cases. Models II, III, and a Gleason score model (model GS) predict sPC that is defined as National Comprehensive Cancer Network (NCCN)-categorized favorable-intermediate risk group or higher (Model II), unfavorable-intermediate risk group or higher (Model III), and GS ≥7 PC (model GS), respectively. The metabolomic panels and predicting models were constructed using logistic regression and Akaike information criterion. Results: The best metabolomic panels from the HILIC column include 25, 27, 28 and 26 metabolites in Models I, II, III, and GS, respectively, with area under the curve (AUC) values ranging between 0.82 and 0.91 in the training cohort and between 0.77 and 0.86 in the validation cohort. The combination of the metabolomic panels and five baseline clinical factors that include serum prostate-specific antigen, age, family history of PC, previously negative biopsy, and abnormal digital rectal examination results significantly increased AUCs (range 0.88–0.91). At 90% sensitivity (validation cohort), 33%, 34%, 41%, and 36% of unnecessary biopsies were avoided in Models I, II, III, and GS, respectively. The above results were successfully validated using LC-MS with the C18 column. Conclusions Urinary metabolomic profiles with baseline clinical factors may accurately predict sPC in men with elevated risk before biopsy.

ObjectiveTo clearly identify the difference between rescue injuries and agonal injuries and to avoid duplicate identifications and misidentifications. MethodsBased on the forensic pathological data of 5 923 cases of death cause identification from 2013 to 2022 in Sun Yat-sen University Forensic Identification Center and Guangzhou Tianhe District Branch of Guangzhou Public Security Bureau， this study retrospectively studied the characteristics of rescue injuries and agonal injuries seen in cause of death identification and their influence on cause of death identification. ResultsAmong all the 5 923 cases， 640 cases were found to have rescue injuries or agonal injuries， and 624 cases received treatment， of which 609 cases were found to have rescue injuries （97.60%）， 44 cases were found to have agonal injuries， and 13 cases were found to have both types of injuries. Among the 640 cases， 441 were male and 199 were female. The age of death was discontinuously distributed from 0 to 95 years old. The leading cause of death was disease， followed by mechanical injury and asphyxia. The main manifestations of rescue injuries were rib and sternum fractures， soft tissue injuries in the prechest area or face， and pericardial rupture. The most common injuries in agonal stage were falling after unconsciousness， inhalation of foreign body in respiratory tract or multiple violent injuries. Among the 640 cases， 19 cases were repeatedly identified， including 15 cases of rescue injuries， 6 cases of agonal injuries， and 2 cases of both types of injuries. Compared with the cases where neither type of injuries was detected， the repeated identification rate of treatment injuries and agonal injuries was significantly increased （χ²=4.04， P=0.044； χ²=43.49， P<0.001）. Among the 640 cases， 11 cases （1.72%） were misidentified as the initial injuries in the first identification， and 13 cases had combined rescue injuries or agonal injuries that were involved in death. ConclusionsBy elucidating the epidemiological characteristics of the two types of injuries， this study proved that the two types of injuries were associated with higher rates of repeated identification and misidentification， which provided a reference for reducing repeated identification and misidentification and improving the accuracy of cause of death identification.

OBJECTIVE: Patients with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection frequently develop central nervous system damage, yet the mechanisms driving this pathology remain unclear. This study investigated the primary pathways and key factors underlying brain tissue damage induced by the SARS-CoV-2 beta variant (lineage B.1.351). METHODS: K18-hACE2 and C57BL/6 mice were intranasally infected with the SARS-CoV-2 beta variant. Viral replication, pathological phenotypes, and brain transcriptomes were analyzed. Gene Ontology (GO) analysis was performed to identify altered pathways. Expression changes of host genes were verified using reverse transcription-quantitative polymerase chain reaction and Western blot. RESULTS: Pathological alterations were observed in the lungs of both mouse strains. However, only K18-hACE2 mice exhibited elevated viral RNA loads and infectious titers in the brain at 3 days post-infection, accompanied by neuropathological injury and weight loss. GO analysis of infected K18-hACE2 brain tissue revealed significant dysregulation of genes associated with innate immunity and antiviral defense responses, including type I interferons, pro-inflammatory cytokines, Toll-like receptor signaling components, and interferon-stimulated genes. Neuroinflammation was evident, alongside activation of apoptotic and pyroptotic pathways. Furthermore, altered neural cell marker expression suggested viral-induced neuroglial activation, resulting in caspase 4 and lipocalin 2 release and disruption of neuronal molecular networks. CONCLUSION These findings elucidate mechanisms of neuropathogenicity associated with the SARS-CoV-2 beta variant and highlight therapeutic targets to mitigate COVID-19-related neurological dysfunction.
Animals ; COVID-19/genetics* ; Mice ; Brain/metabolism* ; Apoptosis ; Mice, Inbred C57BL ; SARS-CoV-2/physiology* ; Pyroptosis ; Gene Expression Profiling ; Transcriptome ; Male ; Female

Objective:To investigate the differences in clinical outcomes of septic children with varying serum zinc levels,and to analyze the relationship between reduced serum zinc levels and organ dysfunction as well as 28-day mortality in septic children.Methods:This study conducted a retrospective analysis of clinical data from pediatric patients diagnosed with sepsis or septic shock in the Department of critical care medicine of the children's Hospital of Soochow University between January 2017 and December 2022.Clinical characteristics,organ dysfunction,and prognosis were compared between two groups:children with low serum zinc levels and those with normal zinc levels.Results:The serum zinc level of septic children within 24 hours of admission was 9.60(5.52,13.80)μmol/L,with 50.54%(94/186)of the children exhibiting low serum zinc levels(<10.07 μmol/L).Compared to the normal serum zinc group,the low serum zinc group had a significantly lower Pediatric Critical Illness Score(PCIS)[(78.71±9.35)vs.(85.12±8.51),P=0.005]and higher 28-day mortality(46.80%vs.14.13%,P<0.001).The low serum zinc group also had a higher proportion of invasive mechanical ventilation(64.89%vs.47.82%,P=0.019),renal replacement therapy(15.59%vs.3.26%,P=0.003),and use of vasoactive drugs(56.38%vs.30.43%,P<0.001).The rate of underlying conditions in the low serum zinc group was significantly higher than that in the normal serum zinc group(57.44%vs.36.95%,P=0.005).Additionally,the low serum zinc group had a higher incidence of disseminated intravascular coagulation(DIC),respiratory failure,acute kidney injury,shock,and multiple organ dysfunction syndrome(MODS)compared to the normal serum zinc group(P<0.05).Serum zinc levels had predictive value for 28-day mortality in septic children(AUC=0.813;95%CI:0.725～0.902;P<0.001).A serum zinc level of less than 6.950 μmol/L predicted the death of septic children with a sensitivity of 0.618 and a specificity of 0.902.Conclusion:Sepsis in children is commonly associated with low serum zinc levels,especially in those with underlying conditions such as hematologic and oncologic disorders.Sepsis patients hypozincemia with a higher incidence of DIC,respiratory failure,acute kidney injury,shock,and MODS.A serum zinc level below 6.95 μmol/L serves as a significant predictor of 28-day mortality in children with severe sepsis.

Objective To analyze the characteristics of intestinal flora of personnel stationed on an island,so as to lay the foundation for maintaining the intestinal microecological balance of personnel stationed on island and provide accurate medical security.Methods Several subjects stationed on an island and several subjects from coastal areas were enrolled by random and sampling method,and their fecal samples were sequenced by 16S rRNA high-throughput sequencing.Diversity and composition of gut microbiota in 2 cohorts of personnel were compared.Results Alpha diversity analysis of intestinal flora showed that the abundance of intestinal flora in subjects stationed on the island was significantly higher than that of subjects from coastal areas.Beta diversity analysis indicated significant differences in the composition of intestinal microbial communities between the subjects stationed on the island and those from coastal areas(P=0.001).The abundance of the Bacteroidota in the intestinal tract of subjects stationed on the island was significantly lower than that of subjects from coastal areas(30.8%vs 48.3%,P＜0.001),while the abundance of the Proteobacteria was significantly higher than that of subjects from coastal areas(28.3%vs 10.2%,P＜0.001).After multiple hypothesis testing correction,it was found that the abundance of the Bacteroides,Roseburia,Alistipes,and Parabacteroides in the intestines of subjects stationed on the island decreased significantly,while the abundance of the Prevotella,Escherichia-Shigella,Citrobacter,and Eubacterium_coprostanoligenes increased significantly.Conclusion The special environment of islands affects the characteristics of intestinal flora of personnel,and the intestinal microecological health needs precise maintenance.

Objective To explore the mechanism of long non-coding RNA fibroblast activation inhibitory factor 1(FAIF1)regulates the proliferation,activation,and fibrosis of human cardiac fibroblasts induced by advanced glycation end products(AGEs).Methods Human cardiac fibroblasts were assigned to control group,AGE group,FAIF1 recombinant lentivirus(Lv-FAIF1)+AGE group or control lentivirus(Lv control)+AGE group.The expression levels of miRNA-424-5p,FAIF1,and Smad7 in myocardial fibroblasts induced by AGEs were detected by quantitative polymerase chain reaction(qPCR)and Western blotting.Bioinformatics analysis was used to predict the interactions between miRNA-424-5p,FAIF1,and Smad7;and luciferase reporter assays were used for verification.Cell proliferation activity was measured by cell counting kit 8 assay,the expression and secretion of collagen Ⅰ/Ⅲ were observed by immunofluorescence staining,and the effect of Lv-FAIF1 on cell activation markers α-smooth muscle actin(α-SMA)and migration proteins matrix metalloproteinase 9(MMP9)induced by AGEs was evaluated by qPCR.Results qPCR and Western blotting results showed that AGEs significantly reduced the expression of FAIF1 and Smad7 in myocardial fibroblasts and upregulated the level of miRNA-424-5p(compared with the control group,all P＜0.05).Bioinformatics analysis revealed that the 3'-untranslated region of Smad7 mRNA contained a binding site for the miRNA-424-5p seed sequence"UGCUGCU",and FAIF1 sequence contained 3 identical binding sites.Luciferase assays showed that miRNA-424-5p inhibited the expression of Smad7,while FAIF1 competed with miRNA-424-5p for binding,thereby relieving the inhibitory effect of miRNA-424-5p on Smad7 mRNA.Functional experiments showed that Lv-FAIF1 significantly inhibited AGEs-induced cell proliferation,collagenⅠ/Ⅲ expression and secretion,as well as α-SMA and MMP9 expression(compared with AGE group,all P＜0.01);and it promoted the expression of Smad7(compared with AGE group,P＜0.01).Conclusion miRNA-424-5p can inhibit the expression of Smad7,and FAIF1 effectively suppresses AGEs-induced over-activation of cardiac fibroblasts by regulating the miRNA-424-5p/Smad7 axis,which provides a new molecular target for the prevention and treatment of diabetic cardiomyopathy.

Objective To explore the application value of a novel portable endoscope to perform upper gastrointestinal tract examinations in primary medical units.Methods A total of 532 subjects receiving portable endoscope examination were enrolled for analysis.The primary outcome was the success rate of operation.The secondary outcomes were the operation time,examination results,polyp removal and biopsy pathology results,and the subjective evaluation.Results In 532 cases,2 were withdrawn midway after the endoscope was inserted into the esophagus due to the patients'inability to tolerate the examination.Additionally,6 cases did not undergo examination of the descending part of the duodenum because of serious reactions during the procedure.Ultimately,524 cases successfully completed the upper gastrointestinal examination,and the success rate was 98.5％.The average examination time was(4.7±1.8)min,and the average time for disposal sheath wearing and removing was(4.2±1.4)min.The most common lesions were chronic non-atrophic gastritis(85.1％,451/530),reflux esophagitis(14.7％,78/530)and bile reflux(14.0％,74/530).A total of 10 cases of polyp removal were completed,and the polyp removal rate was 71.4％(10/14).Biopsy pathological diagnosis was completed in 44 cases,and the biopsy rate was 8.3％(44/530).The main discomfort symptoms during the examination were nausea(53.6％,285/532),vomiting(51.1％,272/532),and sore throat(38.5％,205/532),the main discomfort symptoms after the examination were sore throat(27.8％,148/532),nausea(19.5％,104/532),and vomiting(14.7％,78/532).No serious adverse events such as gastrointestinal bleeding,perforation,cardiac or pulmonary complications occurred.Conclusion The novel portable endoscope can safely and effectively complete the diagnosis and treatment of upper gastrointestinal diseases in primary medical units,while saving the decontamination process.However,the incidence of discomfort is high during examinations.Further optimization of the operation methods is needed.

Objective To investigate the detection rate and related influencing factors of gastrointestinal diseases in grass-roots personnel.Methods A total of 481 grass-roots personnel were enrolled and examined by magnetic-controlled capsule endoscopy(MCCE).Multivariate logistic regression were used to analyze the influencing factors of gastrointestinal diseases detected by MCCE.Results All personnel completed MCCE,and gastrointestinal diseases were detected in 154(32.0%)cases,including 106 cases of erosive gastritis,25 cases of chronic atrophic gastritis,17 cases of digestive tract polyp,16 cases of gastric ulcer,5 cases of reflux esophagitis,4 cases of cardia,1 case of duodenitis,and 1 case of enteritis.Gastrointestinal diseases was correlated with special operation posts,long-term tasks within recent 6 months,abdominal distension,belching,nausea and vomiting,diarrhea and other symptoms(all P＜0.05).Multivariate logistic regression analysis showed that the risk factors of gastrointestinal diseases were working in special operation posts,performing long tasks within 6 months,belching,nausea and vomiting symptoms(all P＜0.05).Conclusion The overall detection rate of gastrointestinal diseases(mainly acid related diseases)is relatively high,and its incidence is closely related to working in special operation posts and performing long-term tasks within recent 6 months.Personnel working in special operation posts should be more alert to gastrointestinal diseases.Gastrointestinal symptoms have reference value for the prediction of lesions,but more attention should be paid to the identification of functional gastrointestinal diseases with endoscopy.

Objective To investigate the effects and underlying mechanisms of down-regulating m6A demethylase fat mass and obesity-associated protein(FTO)on proplatelet formation in the MEG-01 megakaryocytic cells.Methods ①MEG-01 cells were treated with 1 nmol/L phorbol myristate acetate(PMA)(treatment group)or DMSO(control group)for 72 h.FTO expression was measured by Western blotting and RT-qPCR.② MEG-01 cells were infected with targeted FTO shRNA(knockdown group,sh-FTO)or negative control shRNA(negative control group,sh-NC)viruses.FTO knockdown group and negative control group MEG-1 cells were treated with 1 nmol/L PMA for 72 h,and the protein and mRNA expression levels of FTO were detected by Western blotting and RT-qPCR.Cell cycle,viability and apoptosis were assessed by propidium iodide(PI)DNA staining,CCK-8 assay and Annexin V-FITC/PI double staining and TUNEL staining.The expression of cleaved Caspase-3 protein was determine by Western blotting.Megakaryocyte maturation was assessed by CD41/CD61 staining.Proplatelet formation was observed under bright field and detected by CD61 immunofluorescence assay.The expression of apoptosis-related molecules(Caspase3,BAD,BAK1,BCL2 and MCL1)was detected by RT-qPCR,and the protein change of BCL2 was further verified by Western blotting.The dataset was screened out from the gene expression omnibus(GEO)database,and then analyzed with University of California,Santa Cruz(UCSC)genome browser to compare the methylation sequencing peaks on BCL2 mRNA,and m6A methylated RNA immunoprecipitation(m6A-RIP)was used to assess the m6A methylation levels of BCL2 target gene mRNAs in MEG-01 megakaryocytes.Then,the changes in the m6A methylation enrichment level of BCL2 mRNA were observed between the sh-NC group and the sh-FTO group.mRNA stability and ribosome profiling assays were performed to assess translational efficiency of target genes.Results ①PMA treatment upregulated the expression of FTO at protein(P＜0.05)and mRNA(P＜0.01)levels.② FTO shRNA resulted in reduced FTO expression at both mRNA and protein levels(P＜0.01).Compared to the negative control group,the FTO knockdown group showed more cells arrested at the G1/S phase[(60.80±1.29)%vs(72.13±1.18)%,P＜0.01],significantly reduced cell viability[(1.17±0.03)%vs(0.69±0.05)%,P＜0.01],increased Annexin V-FITC/PI positive cells[(12.87±0.83)%vs(17.45±1.58)%,P＜0.01],more TUNEL positive cells[(1.03±0.27)%vs(17.49±9.91)%,P＜0.01],enhanced protein level of cleaved Caspase-3(P＜0.01),decreased proportion of CD41/CD61 positive cells[(51.63±1.13)%vs(34.08±0.53)%,P＜0.01],and less proplatelet formation in MEG-01 megakaryocytes[(26.49±6.73)%vs(13.31±5.97)%,P＜0.01].③Compared to sh-NC group,the FTO knockdown group had significantly decreased protein and mRNA expression of anti-apoptotic molecule BCL2(P＜0.01).UCSC GEO sequencing data revealed there were m6A methylation modification sites on BCL2 mRNA,which was verified through m6A-RIP experiment in MEG-01 megakaryocytes.Compared with GAPDH mRNA,BCL2 mRNA exhibited a significantly enriched m6A signal(P＜0.01).Compared to sh-NC group,a significant increase in m6A methylation modification was observed on BCL2 mRNA.BCL2 mRNA stability was significantly decreased,and its translation efficiency significantly was decreased(P＜0.01).Conclusion m6 A demethylase FTO rgulates BCL2 mRNA stability and translation efficiency,thereby promoting proplatelet formation in MEG-01 megakaryocytes.

Objective To construct a model that can predict the risk of diagnosing ABO-blood group sys-tem hemolytic disease of the newborn(ABO-HDN)and to verify its effectiveness.Methods A total of 446 children with neonatal hyperbilirubinemia who met the inclusion criteria and were first diagnosed in this hos-pital from January 2022 to March 2023 were selected as the modeling group,and were divided into the develo-ping group(200 cases)and the non-developing group(246 cases)according to whether ABO-HDN was diag-nosed.Totally 17 potential influencing factors were included for univariate analysis and multi-factor analysis,and independent risk factors were included in R software to establish a Nomogram model to predict the risk of ABO-HDN.Another 105 cases of neonatal hyperbilirubinemia in the hospital from April to September 2023 were selected as the verification group.Results In the modeling group,Logistic regression analysis showed that maternal pregnancy number,prenatal serum titer,hemoglobin level,white blood cell count,creatine ki-nase level and neonatal Apgar 1 min score were all independent risk factors for ABO-HDN(P＜0.05).Multi-variate Logistic regression analysis showed that the area under receiver operating characteristic(ROC)curve of the modeling group was 0.819(95％CI:0.779-0.859),sensitivity was 0.655,specificity was 0.878.In the verification group,the area under ROC curve was 0.867(95％CI:0.800-0.933),the sensitivity was 0.803,and the specificity was 0.773.Conclusion The established predictive model scoring system can effec-tively predict the risk of ABO-HDN.